NucGreen Dead 488 ReadyProbes Reagent Protocol for Fixed Cells

image of Molecular Probes® NucGreen® Dead 488 ReadyProbes® Reagent

Nuclear stain for fixed cells

NucGreen Dead 488 ReadyProbes Reagent emits bright green fluorescence when bound to DNA and is suitable for staining nuclei in fixed-cell preparations and tissue sections. NucGreen Dead 488 reagent may also be used to measure viability and to measure cytotoxicity in kinetic live-cell assays.

This protocol can be used for:

Nucleic acid (nuclear) staining in fluorescence microscopy

This protocol should not be used for:

Flow cytometry

You will need the following for this protocol:

Cells growing in culture
NucGreen Dead 488 ReadyProbes Reagent (Cat. No. R37109)
Fluorescence microscope with FITC or GFP filters

Protocol

Labeling fixed cells

First, fix and permeabilize cultured cells with a protocol appropriate for your sample.

1. Wash the cells 1–3 times in PBS as needed.

2. Add two drops of NucGreen Dead 488 Reagent per milliliter of medium.

3. Incubate for 5–15 minutes, protected from light.

4. Remove the stain solution.

5. Wash the cells 2–3 times in PBS.

6. Image the cells.

Spectral information and storage

	NucGreen Dead 488
Excitation/Emission (nm)	504/523
Standard filter set	FITC or GFP
EVOS Light Cube	FITC
Storage conditions	Room temperature

Protocol tips

In some cases, more or fewer drops may be needed to achieve optimal staining intensity.
Image quality may be improved by replacing the culture medium with Live Cell Imaging Solution (Cat. No. A14291DJ).

HeLa cells stained with NucGreen Dead 488 ReadyProbes reagent, indicating loss of plasma membrane integrity.